The Evolution of a Female Genital Trait Widely Distributed in the Lepidoptera: Comparative Evidence for an Effect of Sexual Coevolution Vı´ctor Sa´nchez1, Blanca Estela Herna´ndez-Ban˜ os2, Carlos Cordero3* 1 Posgrado en Ciencias Biolo´gicas, Instituto de Ecologı´a, Universidad Nacional Auto´noma de Me´xico, Mexico City, Distrito Federal, Mexico, 2 Departamento de Biologı´a Evolutiva, Facultad de Ciencias, Universidad Nacional Auto´noma de Me´xico, Mexico City, Distrito Federal, Mexico, 3 Instituto de Ecologı´a, Universidad Nacional Auto´noma de Me´xico, Mexico City, Distrito Federal, Mexico Abstract Background: Sexual coevolution is considered responsible for the evolution of many male genital traits, but its effect on female genital morphology is poorly understood. In many lepidopterans, females become temporarily unreceptive after mating and the length of this refractory period is inversely related to the amount of spermatophore remaining in their genital tracts. Sperm competition can select for males that delay female remating by transferring spermatophores with thick spermatophore envelopes that take more time to be broken. These envelopes could select for signa, sclerotized sharp structures located within the female genital tract, that are used for breaking spermatophores. Thus, this hypothesis predicts that thick spermatophore envelopes and signa evolve in polyandrous species, and that these adaptations are lost when monandry evolves subsequently. Here we test the expected associations between female mating pattern and presence/ absence of signa, and review the scant information available on the thickness of spermatophore envelopes. Methodology/Principal Findings: We made a literature review and found information on female mating pattern (monandry/polyandry), presence/absence of signa and phylogenetic position for 37 taxa. We built a phylogenetic supertree for these taxa, mapped both traits on it, and tested for the predicted association by using Pagel’s test for correlated evolution. We found that, as predicted by our hypothesis, monandry evolved eight times and in five of them signa were lost; preliminary evidence suggests that at least in two of the three exceptions males imposed monandry on females by means of specially thick spermatophore envelopes. Previously published data on six genera of Papilionidae is in agreement with the predicted associations between mating pattern and the characteristics of spermatophore envelopes and signa. Conclusions/Significance: Our results support the hypothesis that signa are a product of sexually antagonistic coevolution with spermatophore envelopes. Citation: Sa´nchez V, Herna´ndez-Ban˜os BE, Cordero C (2011) The Evolution of a Female Genital Trait Widely Distributed in the Lepidoptera: Comparative Evidence for an Effect of Sexual Coevolution. PLoS ONE 6(8): e22642. doi:10.1371/journal.pone.0022642 Editor: Daniel Ortiz-Barrientos, The University of Queensland, St. Lucia, Australia Received January 7, 2011; Accepted July 3, 2011; Published August 17, 2011 Copyright: ß 2011 Sa´nchez et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Funding: VS was supported by the Posgrado en Ciencias Biolo´gicas (Universidad Nacional Auto´noma de Me´xico) and a Consejo Nacional de Ciencia y Tecnologı´a scholarship. Programa de Apoyo a Proyectos de Investigacio´n e Inovacio´n Tecnolo´gica/Universidad Nacional Auto´noma de Me´xico (IN223508) and Programa de Apoyo para la Superacio´n del Personal Acade´mico/Direccio´n General de Asuntos del Personal Acade´mico (Universidad Nacional Auto´noma de Me´xico) provided funds to CC, and Consejo Nacional de Ciencia y Tecnologı´a and Programa de Apoyo para la Superacio´n del Personal Acade´mico/Direccio´n General de Asuntos del Personal Acade´mico (Universidad NacionalAuto´noma de Me´xico) to BE. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Competing Interests: The authors have declared that no competing interests exist. * E-mail: [email protected] Introduction morphologically simpler and uniform in several taxa [2,14] is somewhat paradoxical since sexual coevolution predicts evolu- The convergence and divergence of male and female interests tionary responses in both sexes. It can be argued that evolutionary during sexual interactions generates reciprocal selection pressures responses in females are more difficult to detect because they occur that can result in the development of male and female co- at the level of the nervous and endocrine systems [2,3,9], whereas adaptations, a process known as sexual coevolution [1–4]. Depen- male adaptations involve morphological modifications. However, ding on the nature of the selective pressures, sexual coevolution is recent studies indicate that in some groups female morphological driven by mate choice [2,5], sexual conflict (the so-called ‘‘sexually adaptations also have evolved [11,12]. antagonistic coevolution’’; [4,6]) or a mixture of both [7]. Here, we present evidence supporting a sexually antagonistic Empirical evidence supports the hypothesis that sexual coevolution coevolution hypothesis for the evolution of female genital is responsible for the evolution of many male genital traits sclerotized structures called signa, present in many species of ([2,3,6,8–12], but see [13]). As predicted by this hypothesis, in Lepidoptera [15]. Signa are located on the inner wall of the corpus general male genitalia are complex organs that evolve rapidly and bursa, a sac-like organ in which males deposit a spermatophore divergently [2,8,10]. However, the fact that female genitalia are during copulation (Figure 1), and their main function is to break PLoS ONE | www.plosone.org 1 August 2011 | Volume 6 | Issue 8 | e22642 Sexual Coevolution of a Female Genital Trait off the external wall of the spermatophore, thus allowing females length of the period of female sexual refractoriness. The expected access to the resources contained in it [16,17]. Our hypothesis correlations exist in several polyandrous Lepidoptera [15,23,25]. proposes the following sequence of evolutionary steps (Figure 2) (3) Sperm competition generated by polyandry selects for males [15]: (1) Polyandry evolves, possibly to increase the acquisition of that produce spermatophore envelopes more difficult to break, resources contained in spermatophores such as nutrients, hor- thus increasing the lengths of female refractory period and time to mone-like substances, etc. [18–22]. Available data indicates that remating [15,23]. (4) Since the optimal female refractory period is polyandry is widespread in Lepidoptera [2,21,23] and the (also expected to be shorter for females than for their mates (for widespread) taxonomic distribution of polyandry in insects [2,24] example, females may remate to replenish spermatophore suggests that this mating pattern could be plesiomorphic in resources or to ‘‘renew’’ sperm stores when they still have viable Lepidoptera. (2) Polyandrous females evolve an inverse relation- sperm from the previous male), spermatophore envelopes difficult ship between their sexual receptivity and the amount of to break favor the evolution of signa as female devices that increase spermatophore remaining in their corpus bursa to optimize the the rate at which envelopes are torn open, thus moving the rate of balance between replenishment of sperm and spermatophore recovery of sexual receptivity back to the female’s optimum. The resources and remating costs (such as decreased time for foraging process described in (3) and (4) could continue through time and egg laying, predation risk, etc.). This results in a positive (Figure 2) [15]. Therefore, this hypothesis predicts that (a) signa relationship between amount of spermatophore transferred and evolve in polyandrous species, and that (b) if subsequently Figure 1. A sampler of the morphological diversity of signa in female Lepidoptera. Each signum is indicated by an ‘‘S’’. (A) Callophrys xami (Lycaenidae): signa are a pair of thin thorns. (B) Erbessa priverna: (Notodontidae): signum is a plate covered by small thorns. (C) Pyrisitia nise (Pieridae): signum is a strong structure covered by thick spines of different lengths. (D) Ephialtias draconis (Notodontidae): signum is a long, narrow, concave structure with thin spines along the margins of its internal surface. In (A), (C) and (D) the signa are observed through the wall of the corpus bursae, whereas in (B) the corpus bursae was opened and two spermatophores removed. In (B) several deciduous cornuti shed from the male endophallus are attached to the corpus bursae wall, and in (C) there are spermatophore remains within the corpus bursae. db: ductus bursae. Photographs are at different scales. doi:10.1371/journal.pone.0022642.g001 PLoS ONE | www.plosone.org 2 August 2011 | Volume 6 | Issue 8 | e22642 Sexual Coevolution of a Female Genital Trait consistent with current knowledge on Lepidoptera phylogeny [30,31]. Polyandry and presence of signa are plesiomorphic for the taxa analyzed (Figure 3). In agreement with our sexual coevolution hypothesis, only 33.3% of monandrous taxa have signa (3/9) in comparison with 93% of polyandrous taxa (27/29) (Fisher’s exact probability test, P,0.0007). Pagel’s test detected a significant association between female mating pattern and presence/absence